瓜伊马斯深海热液口古菌分布及多样性研究

摘要

Due to the dramatic gradients of temperature (up to above 400℃ ) ,pH and chemicals, deep-sea hydrothermal vents have been the focus for studies of the origin and evolution of life on earth and the subseafloor hydrothermal living system is ideal for exploring the origin and evolution of deep-sea life. In this study, the diversity of archaea was investigated at high temperature sediments and chimney from Guaymas hydrothermal vent using cultured-independent technique. Environmental total DNA were directly extracted and used for the quantification and construction of archaea 16S rRNA gene libraries. A total of 79 and 80 clean sequences were obtained from the sediment and chimney samples. Quantitative PCR analysis revealed that the amounts of archaea present in the sediment and chimney were about 1.47×109 copies/g ( wet weight) and 5. 29 × 108 copies/g ( wet weight), respectively. All the clone libraries had the coverage C value more than 80%, the archaea diversity of sediment sample was higher than the chimney. The compositions of archaea community were different between two samples, yet most of archaea belonged to the thermophilic/hyperthermophilic microorganism because of the high temperature environment. In the sediments, most of archaea came from Crenarchaeota domain and the two most dominant groups were the Miscellaneous Crenarchaeota Group ( MCG,43. 04% )and Hot Water Crenarchaeota Group Ⅰ ( HWCG Ⅰ , 20. 25% ) confirmed by archaeal 16S rRNA gene components. The remaining sequences from the sediment sample were related to members of Thermoprotei ( 7. 59%) , Korarchaeota ( 1. 26%) , Methanosarcinales (1. 26%) , Marine Benthic Group D/Deep Hydrothermal Vent Euryarchaeota ( MBGD/DHVE 1, 8. 86%) , DHVE 9 ( 5. 06%) , Miscellaneous Euryarchaeota Group ( MEG, 2. 53%). While the Euryarchaeota domain dominated the chimney sample, in which Thermococcales (47.5%), Marine Benthic Group D ( MBGD )/Deep Hydrothermal Vent Euryarchaeota 1 ( DHVE 1) ( MBGD/DHVE 1,30% ) were the main components. Phylotypes ascribing to DHVE 2 (6. 25% ) ,DHVE 9 ( 12. 5% ) , Methanococcales ( 1. 25% ) and Hot Water Crenarchaeota Group Ⅰ ( HWCG Ⅰ , 1.25%) were also indemnified from the chimney sample. Moreover, we found a new subgroup of MCG, named MCG-H, which might be a high-temperature specific group of MCG. Therefore, this study will shed some lights on understanding the physiological functions and evolution of archaea.%深海热液口由于剧烈的温度(可达到400℃以上)、pH和化学梯度,成为地球生命起源和演化研究的焦点,海底热液口生命系统为人类提供了一个探索深部生命及生命起源和进化的窗口.本文主要开展了瓜伊马斯深海热液口高温环境下沉积物和烟囱样品中古菌多样性的研究.采用实时荧光定量PCR首先对样品中古菌进行了定量,发现沉积物和烟囱两个样品都含有高丰度的古菌,沉积物和烟囱中古菌分别达到1.47×109拷贝数/g和5.29× 108拷贝数/g(湿重).通过核糖体小亚基16S rRNA基因的PCR扩增,文库构建和系统发育分析,发现沉积物和烟囱样品中的古菌类群多数是嗜热和超嗜热微生物,但两个样品中的古菌群落组成差异很大:沉积物以泉古菌门为主,优势菌群为Miscellaneous Crenarchaeota Group (MCG)和Hot Water Crenarchaeota Group Ⅰ(HWCG Ⅰ)；烟囱样品则主要是广古菌门类群,其中以Thermococcales,Marine Benthic Group D (MBGD)/Deep Hydrothermal Vent Euryarchaeota 1(DHVE 1)为主.在沉积物样品中发现了一个新的MCG类群,命名为MCG-H,可能是MCG的高温独特类群.研究揭示,在瓜伊马斯深海热液口烟囱和沉积物中都包含有丰富的古菌类群包括未被发现的新类群,对瓜伊马斯深海热液口古菌分布和多样性的研究将为进一步深入的古菌生理功能和演化的研究提供基础.